LOCUS       sequence-407159-        8833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for making transgenic fish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8833)
  AUTHORS   Wittamer V, Bertrand JY, Gutschow PW, Traver D
  TITLE     Characterization of the mononuclear phagocyte system in zebrafish.
  JOURNAL   Blood. 2011 Jun 30;117(26):7126-35. doi: 
            10.1182/blood-2010-11-321448. Epub 2011 Mar 15.
  PUBMED    21406720
REFERENCE   2  (bases 1 to 8833)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1182/blood-2010-11-321448"; journalName: "Blood"; date: 
            "2011-06-30- 30"; volume: "117"; issue: "26"; pages: "7126-35"
COMMENT     Sequence Label: mhc2dab:GFP-LT
FEATURES             Location/Qualifiers
     source          1..8833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(1843..2005,2006..2017)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     CDS             2029..2040
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2050..2070)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2050..2068)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2112..2128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2112..2128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2125..2147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2136..2152
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2160..2166,2167..2184,2185..2190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2205..2226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2343..2360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2514..3102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2594..2613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3273..4064,4065..4133))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3896..3915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4134..4238)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4264..4719)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4401..4422)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4613..4632
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4846..4868
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4860..4877
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4861..4877
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4910..4929
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4910..4928
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(4956..4967,4968..5155)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     polyA_signal    5594..5728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5631..5650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5685..5704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5738..6451)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5778..5798)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     6393..6421
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 tatttccttt tatgtaatta atttatttta tttcacagta aatataagtg accaatgtaa
       61 aaaaaaaaat gtcagtatat atgtggaata atctttaact aaatatttaa ctatatttag
      121 gtttatttta tagaaacaga tgaaactgtt aattgtaaca ctgtttgcaa aatatttagg
      181 tcatattttt gtcagttctt ctaactaaaa atactgttta atttaattct atttatttta
      241 gtttaattaa gtttaccaaa gccttgaaat atatcttttt tttattgaat acaaatgatt
      301 taatacaaat catatttgtt tgtaacacac tcaatttaaa ttatgctaaa cagatgtcgt
      361 taataataat aaatatgttc atatttcaaa taatcatgtc tactcattgc ttaaagattt
      421 gtttgtaaca attttcactg cccacacagc tgacattgtg attgtgatat gtaaaaagtt
      481 tttaatgact aaaaagcaga cacaatagtt tcaatttaat tatttatttc taaaggtttg
      541 attattcagt atcaaacaac tgtgcaaaaa aaaaaagact gaataaaaga tcttaaagta
      601 ccactattta aagttgcatt ataaaaccac actcgcatta cctgatttaa aattcaatga
      661 aaatgcataa atgatcaaag catgaaaaaa ttacaattaa attaagcttt gcaattaagt
      721 ttaaatttag cataaatttc aagacccttt tgtctttgct tatcataaga taaaaccaga
      781 acacaaaata acgtgtagaa ttgtcacaag tcacaaaaat aagacatatc cggaagcaaa
      841 aatgatttaa tggaacaatt aaatagtgca tatatgtgtg tttatgtgac tgtttacttc
      901 agaggacaag tggatttact ggcaaaaaat atcaacaaaa gttttgtgaa tatattttat
      961 gtgtctttta gttaaggcaa ataactgaaa attttttttt tacaggatca agtttatgga
     1021 gaagaaaata taattcatct aacctaaaag ctatggaatc taaatgtcct gagaaggata
     1081 tagtcccaaa cctgtatgaa tgcctaataa tttgtatggt aactttgatc atcaaaactt
     1141 tcgattatta ttgtcacaca cacacacaca cacacacaca cacacacaca cacacacaca
     1201 cacacacaca cacacacact gtctttttcc ttataccacc atattggatt gtttaaaaat
     1261 tatatccttt atatttaaat gaagggcgaa gcagtggtac agtaggtagt gctgtcgcct
     1321 cacagcaaga aggtcgctgg gtcgctggtt cgagcctcag ctcagttggc gtttctgtga
     1381 agagtttgca tgttcttcct gcgttcgtgt tggtttcctc caggtactcc ggttttcccc
     1441 acagtccaaa gacatgcggt acgcggccgc caccgcggcc catctggcct gtgtttcaga
     1501 caccagggag tctctgctca cgtttcctgc tatttgcagc ctctctatca agactaatac
     1561 acctcttccc gcatcggctg cctgtgagag gcttttcagc actgcaggat tgcttttcag
     1621 ccccaaaaga gctaggcttg acactaacaa ttttgagaat cagcttctac tgaagttaaa
     1681 tctgaggttt tacaactttg agtagcgtgt actggcatta gattgtctgt cttatagttt
     1741 gataattaaa tacaaacagt tctaaagcag gataaaacct tgtatgcatt tcatttaatg
     1801 ttttttgaga ttaaaagctt aaacaagaat ctctagtttt ctttcttgct tttactttta
     1861 cttccttaat actcaagtac aattttaatg gagtactttt ttacttttac tcaagtaaga
     1921 ttctagccag atacttttac ttttaattga gtaaaatttt ccctaagtac ttgtactttc
     1981 acttgagtaa aatttttgag tactttttac acctctgtca agaactcctg ggagctccag
     2041 cttttgttcc ctttagtgag ggttaattgc gcgcattacc ctgttatccc tacgcgcttg
     2101 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     2161 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     2221 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     2281 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2341 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2401 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2461 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2521 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2581 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2641 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2701 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2761 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2821 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2881 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2941 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3001 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3061 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3121 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3181 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3241 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3301 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3361 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3421 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3481 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3541 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3601 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3661 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3721 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3781 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3841 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3901 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3961 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4021 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4081 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4141 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4201 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4261 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     4321 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     4381 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4441 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4501 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4561 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4621 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4681 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     4741 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     4801 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     4861 gtaaaacgac ggccagtgag cgcgtaggga taacagggta atgcgcgcgt aatacgactc
     4921 actatagggc gaattgggta ccgggggtca agaaccagag gtgtaaagta cttgagtaat
     4981 tttacttgat tactgtactt aagtattatt tttggggatt tttactttac ttgagtacaa
     5041 ttaaaaatca atacttttac ttttacttaa ttacattttt ttagaaaaaa aagtactttt
     5101 tactccttac aattttattt acagtcaaaa agtacttatt ttttggagat cacttcattc
     5161 tattttccct tgctattacc aaaccaattg aattgcgctg atgcccagtt taatttaaat
     5221 gttatttatt ctgcctatga aaatcgtttt cacattatat gaaattggtc agacatgttc
     5281 attggtcctt tggaagtgac gtcatgtcac atctattacc acaatgcaca gcaccttgac
     5341 ctggaaatta gggaaattat aacagtcaat cagtggaaga aaatggagga agtatgtgat
     5401 tcatcagcag ctgcgagcag cacagtccaa aatcagccac aggatcaaga gcacccgtgg
     5461 ccgtatcttc gcagatctga tctagaggat cataatcagc cataccacat ttgtagaggt
     5521 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     5581 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5641 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5701 catcaatgta tcttatcatg tctgaatcct cgagttattt gtatagttca tccatgccat
     5761 gtgtaatccc agcagctgtt acaaactcaa gaaggaccat gtggtctctc ttttcgttgg
     5821 gatctttcga aagggcagat tgtgtggaca ggtaatggtt gtctggtaaa aggacagggc
     5881 catcgccaat tggagtattt tgttgataat ggtctgctag ttgaacgctt ccatcttcaa
     5941 tgttgtgtct aattttgaag ttaactttga ttccattctt ttgtttgtct gccatgatgt
     6001 atacattgtg tgagttatag ttgtattcca atttgtgtcc aagaatgttt ccatcttctt
     6061 taaaatcaat accttttaac tcgattctat taacaagggt atcaccttca aacttgactt
     6121 cagcacgtgt cttgtagttc ccgtcatctt tgaaaaatat agttctttcc tgtacataac
     6181 cttcgggcat ggcactcttg aaaaagtcat gccgtttcat atgatctggg tatcttgaaa
     6241 agcattgaac accataagtg agagtagtga caagtgttgg ccatggaaca ggtagttttc
     6301 cagtagtgca aataaattta agggtaagtt ttccgtatgt tgcatcacct tcaccctctc
     6361 cactgacaga aaatttgtgc ccattaacat caccatctaa ttcaacaaga attgggacaa
     6421 ctccagtgaa aagttcttct cctttactca tggtgaattc ctgcagcccg ggggatccga
     6481 gtctttgaat gtgtcaaatg aagaactttc cctcagtgtc ttcttcttgt ggtggatttc
     6541 taaataatct gaccaatcag cttactccca cataagtgtt gctaggtcaa aaactgcttt
     6601 catttttctg ttttcacaag acaattactg tgagtgcatt aacacatgta taaccatagc
     6661 ggttgtgtac actacacaaa aaatgataaa atgtaagaat atttattagt tgtgtctgtt
     6721 ttctaaatta agaacatgtg tttatattag attagtttag ttatattttt tatagaaaca
     6781 gatgaaactg tttatagttg tgacagtttg ctatatattt atttggtaac atttttgtaa
     6841 attacaattt tttattttat ttaactaaag tcttgaaatg taatttaatt ttttaacatc
     6901 aatatattga aataatgtca atatatttat ttgcaaaaaa aaagatttta aaatttattt
     6961 tattttatta tttcatttta gttaagccta atttataatt ttattgtatt ttatatttta
     7021 ttttattaac gtttagtgtg ttattttatt taactttaat ttgatatatt actttattgt
     7081 tttgttttat ttgatttgat tttactttat tttgctttgg tttttgtttt aatttattat
     7141 tttttatttt attcagtcaa tatattttat tcagtattgt ttgagtattc tattagtttt
     7201 attctattat ttcaatttat tttagtttac tttagtttat tattttatta catacatttt
     7261 attttattaa ttgtattaac gtttggtata ttattttagt ttactttatt attttacttt
     7321 attttaatga ctgattttat tttactttat tttgctttga ttttagtttg attttatttt
     7381 acttattgtt ttattttatt tagtcaatat attgttgttc aatatatatt taattattgt
     7441 atatatttta ttattgtatt ttatgttagt ttattttaaa ttaatttatt gttttacttt
     7501 attttgatgt cttattttat ttttctttat tgttttattc atattattag ttatttttat
     7561 tttattaatt taatatttta ttcagtcaat atatttatta tatataattt attcatttat
     7621 taataatata atttcttttt gtattttagt ttagttaagg ctattatttt atttaattat
     7681 tttaattgtt ggctttgctt tgttttatgt tatttttttt aatttcttat tatttatttg
     7741 tttataactt attatttatg tctgtataat attttcgtta attcttcaaa tgttttgttt
     7801 tgttttattt tagttataaa aatgtataaa aattttgttt tttaataaat attttgtcag
     7861 tcaatatatg tttattcatt tttttatttt tttattttat gatatggttt tcttttagct
     7921 tattttcttt tattatttta ttttagatta ttttataata ttaattttat ttaagcttat
     7981 tttattttaa ttattgttta ttttaattta atatttttat cattttattt tagtttactg
     8041 ttattattat tatttttttt tattatttat atgttttttg tttgttttat tatttatgtc
     8101 tgtgtggtat tttttttaaa tttccaaaag tattgttttg tttagtttta ttttagttta
     8161 tcttaaaaat ttataaaaaa aacgattttt aataaatatt ttgtcaatct ttatgtttat
     8221 ttaatatttt ttattattta gttttattat ttaatttgaa tttttttatt tctttatttt
     8281 attaagttta gtttattgtt tattttattt taatttaatt atatatttta ttttagtgca
     8341 tcattttagt ttattatttt atttttattt tattttattt ttttaatttt ttttatttta
     8401 ttttgtttgt tttattttag tttagtttaa aatttaattt aaaaaaaaat atttttgtca
     8461 agcaatatat ccattcatta attttccttc ggcttagtcc ctatatttca gaacctccaa
     8521 ctattccagc atatgtttta tgcggtggat gcccttccag ccgtaaccct gttctgggaa
     8581 acatctatac actcgtacat acactaagtt tgatttagct cattcaattc acttttagca
     8641 catgtttgaa ctgtggggga aaccggagca cccagaggaa atcaatgcaa acatgaggag
     8701 aacatgcaac tccacgcaga aatgccaaca ggctcagcca ggtgagcgac agtgctaacc
     8761 actgagccat tttttttcta ttagtttatt attttatttg agattatttt taatttattt
     8821 tatttgtgtt cat
//
